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Dynamics of human DNA topoisomerases II and IIß in living cells

¹ Department of Clinical Chemistry, Medizinische Poliklinik, University of Würzburg, D-97070 Würzburg, Germany
² Department of Molecular and Structural Biology, University of Aarhus, DK-8200 Aarhus-C, Denmark
³ Department of Clincal Genetics, University of Aarhus, DK-8200 Aarhus-C, Denmark
⁴ Department of Cell and Developmental Biology, Biocenter, University of Würzburg, D-97074 Würzburg, Germany
⁵ Cancercytogenetics Laboratory, Aarhus Amtssygehus, Aarhus University Hospital, DK-8000 Aarhus C, Denmark

Address correspondence to Christian Mielke, Dept. of Clinical Chemistry, Medizinische Poliklinik, University of Würzburg, Klinikstrasse 6-8, D-97070 Würzburg, Germany. Tel.: 49-931-201-7008. Fax: 49-931-201-7098. E-mail: christian.mielke{at}mail.uni-wuerzburg.de

DNA topoisomerase (topo) II catalyses topological genomic changes essential for many DNA metabolic processes. It is also regarded as a structural component of the nuclear matrix in interphase and the mitotic chromosome scaffold. Mammals have two isoforms ( and ß) with similar properties in vitro. Here, we investigated their properties in living and proliferating cells, stably expressing biofluorescent chimera of the human isozymes. Topo II and IIß behaved similarly in interphase but differently in mitosis, where only topo II was chromosome associated to a major part. During interphase, both isozymes joined in nucleolar reassembly and accumulated in nucleoli, which seemed not to involve catalytic DNA turnover because treatment with teniposide (stabilizing covalent catalytic DNA intermediates of topo II) relocated the bulk of the enzymes from the nucleoli to nucleoplasmic granules. Photobleaching revealed that the entire complement of both isozymes was completely mobile and free to exchange between nuclear subcompartments in interphase. In chromosomes, topo II was also completely mobile and had a uniform distribution. However, hypotonic cell lysis triggered an axial pattern. These observations suggest that topo II is not an immobile, structural component of the chromosomal scaffold or the interphase karyoskeleton, but rather a dynamic interaction partner of such structures.

Key Words: DNA topoisomerase II; cell cycle; nucleolus; nuclear matrix; chromosomal scaffold

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